FGF-23 transgenic mice demonstrate hypophosphatemic rickets with reduced expression of sodium phosphate cotransporter type IIa

Fibroblast growth factor (FGF)-23 was identified as a causative factor of tumor-induced osteomalacia and also as a responsible gene for autosomal dominant hypophosphatemic rickets. To clarify the pathophysiological roles of FGF-23 in these diseases, we generated its transgenic mice. The transgenic mice expressing human FGF-23 reproduced the common clinical features of these diseases such as hypophosphatemia probably due to increased renal phosphate wasting, inappropriately low serum 1,25-dihydroxyvitamin D level, and rachitic bone. The renal phosphate wasting in the transgenic mice was accompanied by the reduced expression of sodium phosphate cotransporter type IIa in renal proximal tubules. These results reinforce the notion that the excessive action of FGF-23 plays a causative role in the development of several hypophosphatemic rickets/osteomalacia.     

Asymmetric larval interactions between introduced and indigenous ladybirds in North America

Understanding the mechanisms that result in the success of introduced species will contribute to predicting future invasions and managing invaded systems. We examined interactions between larvae of two predatory ladybird species recently introduced to North America, Coccinella septempunctata (CS) and Harmonia axyridis (HA), and two indigenous ladybirds, Coccinella transversoguttata (CT) and Hippodamia convergens (HC). By pairing young and old larvae in the laboratory at low and high levels of aphid availability, we assessed the degree of asymmetry in intraguild predation (IGP), the strength of competitive effects on growth and development of larvae escaping predation, and the nature of attack and escape behavior among the species. Interactions were generally asymmetric, with larvae of introduced species acting most frequently as intraguild predators and larvae of indigenous species serving most frequently as intraguild prey (the two Coccinella spp., however, preyed on each other at similar rates). Because they were especially aggressive and because other larvae were least successful in escaping their attacks, larvae of HA had stronger negative effects on larvae of the two indigenous species than did larvae of CS. Such negative effects, expressed most strongly when aphid availability was low, were especially adverse for the smaller of the two indigenous species, HC. In general, older larvae interacted with each other more strongly than young larvae did, and older larvae had especially strong negative effects on young larvae when interactions occurred between age classes. Our results suggest that HA more than CS may represent a threat to indigenous ladybirds as an intraguild predator, and that IGP in turn may play a stronger role for HA than for C. septempunctata in promoting the successful invasion of North America.     

Contractile and relaxant effects of jellyfish toxin on the vascular and intestinal tissues

A toxic component (pCrTX) of jellyfish Carybdea rastonii (10(-8)-10(-6)g/ml) caused a contraction in both rat aorta and guinea-pig taenia coli which was partially inhibited by indomethacin or aspirin. pCrTX (10(-7)-10(-6)g/ml) relaxed the norepinephrine-induced contraction in rat aorta which was inhibited by removing endothelium or by adding methylene blue. These results suggest that a portion of the pCrTX-induced contraction is due to release of prostaglandin(s) and that the pCrTX-induced relaxation is due to release of an endothelium-derived relaxing factor.     

Intracellular Ca2+-calmodulin system involved in the palytoxin-induced K+ release from rabbit erythrocytes

Palytoxin (PTX) caused K+ release from rabbit erythrocytes which was dependent on the concentrations of extracellular Ca2+ and PTX. In a Ca2+-free solution, PTX still caused a slow K+ release. An intracellular Ca2+ antagonist, TMB-8, an intracellular Ca2+ chelator, quin 2, and calmodulin inhibitors, prenylamine, W-7 and W-5, inhibited the PTX-induced K+ release in a Ca2+-free solution. These results suggest that the PTX-induced K+ release is dependent on the process including intracellular Ca2+ and calmodulin.     

Synthesis and characterization of chimeric particles between epizootic hemorrhagic disease virus and bluetongue virus: functional domains are conserved on the VP3 protein.

A functional assay has been developed to determine the conservative nature of the interacting sites of various structural proteins of orbiviruses by using baculovirus expression vectors. For this investigation, proteins of two serologically related orbiviruses, bluetongue virus (BTV) and the less studied epizootic hemorrhagic disease virus (EHDV), were used to synthesize chimeric particles. The results demonstrate that the inner capsid protein VP3 of EHDV-1 can replace VP3 protein of BTV in formation of the single-shelled corelike particles and the double-shelled viruslike particles. Moreover, we have demonstrated that all three minor core proteins (VP1, VP4, and VP6) can be incorporated into the homologous and chimeric corelike and viruslike particles, indicating that the functional epitopes of the VP3 protein are conserved for the morphological events of the virus. This is the first evidence of assembly of seven structural proteins of the virus by a baculovirus expression system. Confirmation at the molecular level was obtained by determining the EHDV-1 L3 gene nucleic sequence and by comparing it with sequences available for BTV. The analysis revealed a high degree homology between the two proteins: 20% difference, 50% of which is conservative. The consequences for Orbivirus phylogeny and the possibility of gene reassortments are discussed.     

Involvement of the visual change detection process in facilitating perceptual alternation in the bistable image

A bistable image induces one of two perceptual alternatives. When the bistable visual image is continuously viewed, the percept of the image alternates from one possible percept to the other. Perceptual alternation was previously reported to be induced by an exogenous perturbation in the bistable image, and this perturbation was theoretically interpreted to cause neural noise, prompting a transition between two stable perceptual states. However, little is known experimentally about the visual processing of exogenously driven perceptual alternation. Based on the findings of a previous behavioral study (Urakawa et al. in Perception 45:474–482, 2016), the present study hypothesized that the automatic visual change detection process, which is relevant to the detection of a visual change in a sequence of visual events, has an enhancing effect on the induction of perceptual alternation, similar to neural noise. In order to clarify this issue, we developed a novel experimental paradigm in which visual mismatch negativity (vMMN), an electroencephalographic brain response that reflects visual change detection, was evoked while participants continuously viewed the bistable image. In terms of inter-individual differences in neural and behavioral data, we found that enhancements in the peak amplitude of vMMN1, early vMMN at a latency of approximately 150 ms, correlated with increases in the proportion of perceptual alternation across participants. Our results indicate the involvement of automatic visual change detection in the induction of perceptual alternation, similar to neural noise, thereby providing a deeper insight into the neural mechanisms underlying exogenously driven perceptual alternation in the bistable image.     

Identification of bluetongue virus VP6 protein as a nucleic acid-binding protein and the localization of VP6 in virus-infected vertebrate cells.

Recently the insect baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) has been effectively adapted as a highly efficient vector in insect cells for the expression of various genes. A cDNA sequence of RNA segment 9 of bluetongue virus serotype 10 (BTV-10, an orbivirus member of the Reoviridae family) encoding a minor core protein (VP6) has been inserted into the BamHI site of the pAcYM1 transfer vector derived from AcNPV. Spodoptera frugiperda cells were cotransfected with the derived vector in the presence of authentic AcNPV DNA to produce recombinant viruses. These synthesized significant amounts of a protein (representing ca. 50% of the stained cellular protein) similar in size and antigenicity to the authentic BTV VP6. The expressed protein was identified as a nucleic acid-binding protein by using an RNA overlay-protein blot assay. A polyclonal anti-VP6 serum prepared by using the expressed VP6 protein has been used in an immunogold procedure to locate VP6 in BTV-infected mammalian cells. Gold was found to be associated with the matrix of virus inclusion bodies (VIB), with viruslike particles in the VIB, as well as with mature virion particles that were in close proximity to the VIB or were released from cells and adsorbed to cell surfaces. The recombinant virus antigen has also been used to identify antibodies to different BTV serotypes in infected sheep sera, indicating the potential of the expressed protein as a group-reactive antigen for the diagnosis of BTV infections.     

Nuclear transfer of blastomeres expressing EGFP-reporter gene may improve the efficiency of transgenic cattle

The effect of timing of microinjection of DNA constructs on the efficiency of transgenic embryo production and improved efficiency and quality through combining EGFP as a reporter gene with nuclear transfer techniques were examined. From 12 to 24 h after insemination, constructs of pCXNeo-EGFP were microinjected into a pronucleus of bovine IVM-IVF zygotes. Due to the difficulty in visualizing pronuclei, the incidence of successful injection of linear DNA was higher when zygotes were injected between 20 and 24 h, as compared with an early period between 12 and 16 h after insemination. However, developmental competence of DNA-injected zygotes and the EGFP expression rate were not affected by the injection time. A majority of the embryos expressing EGFP signal were mosaic. Following nuclear transfer of blastomeres expressing EGFP, 4.5% of morulae that developed from the NT eggs had a strong EGFP signal in all live blastomeres. In other embryos, EGFP signal had been lost. When cells derived from the EGFP-positive NT morulae were subcultured, all the cells expressed strong EGFP signal at the second passage and demonstrated neomycin resistance. These results show that transient expression of nonintegrated EGFP appears frequently in EGFP-positive bovine embryos and that additional selection of EGFP-positive morulae after nuclear transfer of EGFP-positive blastomeres would facilitate selection of transgenic embryos.